Effects of semen collection methods on sperm quality and metabolite profile in goat seminal plasma: Comparing between artificial vagina and electro-ejaculator techniques.

This study compared two semen collection methods-artificial vagina (AV) and electro-ejaculator (EE)-to evaluate their effects on sperm quality and the metabolomic profile of goat seminal plasma (SP). Semen samples were analyzed for sperm characteristics, and SP was assessed using ultra-performance liquid chromatography-electrospray ionization quadrupole Orbitrap mass spectrometry. Sperm collected via AV showed lower ejaculate volume but significantly higher concentration and motility than EE. Metabolomic profiling revealed 568 and 350 metabolites in positive and negative ion modes, respectively, with 105 and 77 showing significant differential expression. In the negative ion mode, metabolites such as bezafibrate, phosphatidylinositol 16 and 18, and benzothiazoline were down-regulated in AV, while docosahexaenoic acid, arachidonic acid, phosphatidylcholine, rutin, pyruvate, famotidine, and citrate were up-regulated. In the positive ion mode, metabolites including astaxanthin, lapachol, and 3-acetylaconitine were down-regulated, while mannitol, choline, L-carnitine, and indirubin were up-regulated in AV. KEGG pathway enrichment showed 30 significantly altered pathways. This is the first study to assess the impact of AV and EE on the metabolomic profile of goat SP. The identified metabolites serve as promising biomarkers for evaluating sperm quality and assessing reproductive potential, with implications for improving semen collection techniques and conserving genetic resources.